【作者】 王革； 尤明强； 李睿； 胡丽娜； 马维民； 卜培英； 王秉翔；

【Author】 WANG Ge,YOU Ming-qiang,LI Rui,HULi-na,MA Wei-min, BU Pei-ying,WANG Bing-xiang (Lanzhou Institute of Biological Products,Lanzhou 730046,China)

【机构】 兰州生物制品研究所；

【摘要】 对表达炭疽保护性抗原(PA)的重组菌株进行发酵,并纯化、分析目的蛋白PA的生物学特性。试验结果显示重组菌株在LB、TB培养基中发酵,不限制通氧量、pH维持在6.8～7.6细胞株生长所能接受的范围、30℃条件下诱导,可获得可溶性目的蛋白PA;诱导产物经离子交换层析、疏水层析及凝胶过滤层析可溶性目的蛋白PA纯度大于90%;分别以SDS-PAGE、Western blot、免疫双扩散及免疫小鼠后ELISA检测鼠血清中抗-PA抗体水平分析可溶性目的蛋白PA的生物学特性,与天然PA相同。更多还原

【Abstract】 This report describes the production of PA from a recombinant of E.coli strain.And the subsequent fermentation,purification and assay for characterization of the protein product.The protein product in LB、TB medium fermentation and under 30℃induction nonlimiting aeration pH 6.8～7.6 is dissolubility.A purity of 55%～65%was achieved for rPA by diafiltration anion-exchange chromatography,while 80%～90%purity was achieved for rPA by hydrophobic interaction chromatography,and final purity with an additional gel filtration.The final purity of protein product is more than 90%.The purity of the rPA product was characterized by SDS-PAGE,ELISA,Western blot assay. The biological activity of the rPA determined as same as native PA.更多还原